Yiqi Huoxue Compound promotes neuroprotection against ischemic stroke by inhibiting apoptosis through the activation of PI3K/AKT pathway

Yiqi Huoxue Compound (YQHX), a traditional Chinese medicine formulation, has demonstrated therapeutic potential for ischemic stroke (IS). However, its underlying mechanisms are not fully understood. This study aimed to elucidate the neuroprotective effects and mechanisms of YQHX using an integrated strategy encompassing UHPLC-Q-Orbitrap HRMS-based chemical profiling, in vivo pharmacodynamic evaluation, transcriptomics, network analysis, molecular docking, and in vitro validation. Chemical analysis identified 196 constituents in YQHX, mainly terpenoids, saccharides, and polyphenols. In a mouse model of middle cerebral artery occlusion (MCAO), treatment with low, medium, or high doses of YQHX significantly improved neurological function scores, ameliorated cerebral histopathological damage, increased Nissl bodies, and reduced neuronal apoptosis. This was evidenced by decreased TUNEL-positive cells and regulated expression of apoptosis-related proteins (Bax, Bcl-2, cleaved-caspase 3). Transcriptomic analysis of brain tissue highlighted the PI3K-AKT signaling pathway as a key target, which was further supported by molecular docking showing strong binding affinities between core YQHX components and pathway targets. Importantly, YQHX administration upregulated the p-PI3K/PI3K and p-AKT/AKT ratios and attenuated mitochondrial ultrastructural damage in MCAO mice. In vitro, using HT22 cells subjected to oxygen-glucose deprivation/reoxygenation (OGD/R), YQHX treatment suppressed inflammatory cytokine release (TNF-α, IL-6, IL-1β), inhibited apoptosis, and restored the balance of apoptosis-related and PI3K/AKT pathway proteins. Collectively, these results demonstrate that YQHX protects against ischemic brain injury by activating the PI3K/AKT pathway, thereby reducing inflammation and apoptosis. This study provides a mechanistic foundation for the clinical application of YQHX in IS.